INVESTIGATION OF THE EFFECTS OF PREGABALIN ON WOUND HEALING IN L929 FIBROBLAST CELLS

Year 2024, Volume: 1 Issue: 1, 8 - 14, 28.02.2024

Hüseyin Bekmez Hamza Halıcı

https://doi.org/10.5281/zenodo.10712118

Abstract

Aim: Wound healing is a multifaceted, complex process consisting of sequential and interrelated phases including hemostasis/inflammation phase, proliferation phase and remodeling phase. Pregabalin (PGB), a gabapentin derivative, is an anticonvulsant agent with anti-inflammatory and antioxidant properties. Therefore, in this study, we aimed to show the effect of pregabalin on cell viability in L929 fibroblast cells and its effects on fibroblast migration and wound closure during the wound healing process.
Materials And Methods: In this study, the effect of different concentrations of pregabalin on cell viability and proliferation in L929 skin fibroblast cells was investigated using MTT assay. In addition, a scratch wound healing model was established in L929 skin fibroblast cells and the effects of pregabalin concentrations that increase cell proliferation on wound healing in MTT assay were shown. At the end of the experiment, TGF-β1 levels of all groups were measured by ELISA method.
Results: In our studies, it was observed that 100, 50, 25, 10 µM concentrations of pregabalin increased cell proliferation. In the scratch wound healing model, pregabalin at concentrations of 100 and 50 µM showed a significant closure compared to control and other groups. TGF-β1 levels were decreased in groups with good healing scores (50, 25 µM).
Conclusion: Pregabalin has been shown to enhance wound healing in in vitro experiments. This effect needs to be evaluated holistically within the organ system in vivo. There is also a need for experimental and clinical studies to evaluate the wound healing effects and mechanism of pregabalin.

Keywords

Wound Healing, pregabalin, scratch wound assay, L929 cell line

References

• Abou-Khalil, B. W. (2016). Antiepileptic Drugs. Continuum (Minneap Minn), 22(1 Epilepsy), 132-156. https://doi.org/10.1212/CON.0000000000000289
• Abu-Rish, E. Y., Mansour, A. T., Mansour, H. T., Dahabiyeh, L. A., Aleidi, S. M., & Bustanji, Y. (2020). Pregabalin inhibits in vivo and in vitro cytokine secretion and attenuates spleen inflammation in Lipopolysaccharide/Concanavalin A -induced murine models of inflammation. Sci Rep, 10(1), 4007. https://doi.org/10.1038/s41598-020-61006-1
• Arango Duque, G., & Descoteaux, A. (2014). Macrophage cytokines: involvement in immunity and infectious diseases. Front Immunol, 5, 491. https://doi.org/10.3389/fimmu.2014.00491
• Barrientos, S., Stojadinovic, O., Golinko, M. S., Brem, H., & Tomic-Canic, M. (2008). Growth factors and cytokines in wound healing. Wound Repair Regen, 16(5), 585-601. https://doi.org/10.1111/j.1524-475X.2008.00410.x
• Berman, B., Maderal, A., & Raphael, B. (2017). Keloids and Hypertrophic Scars: Pathophysiology, Classification, and Treatment. Dermatol Surg, 43 Suppl 1, S3-S18. https://doi.org/10.1097/DSS.0000000000000819
• Borges, G. A., Elias, S. T., da Silva, S. M., Magalhaes, P. O., Macedo, S. B., Ribeiro, A. P., & Guerra, E. N. (2017). In vitro evaluation of wound healing and antimicrobial potential of ozone therapy. J Craniomaxillofac Surg, 45(3), 364-370. https://doi.org/10.1016/j.jcms.2017.01.005
• Cangul, S., Adiguzel, O., & Tekin, S. (2020). Comparison of Cytotoxicity of Four Different Adhesive Materials Before and After Polymerisation. Oral Health & Preventive Dentistry, 18(1), 43-51. https://doi.org/10.3290/j.ohpd.a43940
• Ceyhan M. , T. E. (2008). Yeni Bir Antikonvülsan Pregabalin. Turkish Journal of Neurology, 14(3), 161-171.
• Cory, G. (2011). Scratch-Wound Assay. Cell Migration: Developmental Methods and Protocols, Second Edition, 769, 25-30. https://doi.org/10.1007/978-1-61779-207-6_2
• Desmouliere, A., Geinoz, A., Gabbiani, F., & Gabbiani, G. (1993). Transforming growth factor-beta 1 induces alpha-smooth muscle actin expression in granulation tissue myofibroblasts and in quiescent and growing cultured fibroblasts. J Cell Biol, 122(1), 103-111. https://doi.org/10.1083/jcb.122.1.103
• El Ayadi, A., Jay, J. W., & Prasai, A. (2020). Current Approaches Targeting the Wound Healing Phases to Attenuate Fibrosis and Scarring. Int J Mol Sci, 21(3). https://doi.org/10.3390/ijms21031105
• Eutamene, H., Coelho, A. M., Theodorou, V., Toulouse, M., Chovet, M., Doherty, A., Fioramonti, J., & Bueno, L. (2000). Antinociceptive effect of pregabalin in septic shock-induced rectal hypersensitivity in rats. J Pharmacol Exp Ther, 295(1), 162-167. https://www.ncbi.nlm.nih.gov/pubmed/10991974
• Everts, P. A., Knape, J. T., Weibrich, G., Schonberger, J. P., Hoffmann, J., Overdevest, E. P., Box, H. A., & van Zundert, A. (2006). Platelet-rich plasma and platelet gel: a review. J Extra Corpor Technol, 38(2), 174-187. https://www.ncbi.nlm.nih.gov/pubmed/16921694
• Fehrenbacher, J. C., Taylor, C. P., & Vasko, M. R. (2003). Pregabalin and gabapentin reduce release of substance P and CGRP from rat spinal tissues only after inflammation or activation of protein kinase C. Pain, 105(1-2), 133-141. https://doi.org/10.1016/s0304-3959(03)00173-8
• Gauglitz, G. G., Korting, H. C., Pavicic, T., Ruzicka, T., & Jeschke, M. G. (2011). Hypertrophic scarring and keloids: pathomechanisms and current and emerging treatment strategies. Mol Med, 17(1-2), 113-125. https://doi.org/10.2119/molmed.2009.00153
• Hecker, A., Schellnegger, M., Hofmann, E., Luze, H., Nischwitz, S. P., Kamolz, L. P., & Kotzbeck, P. (2022). The impact of resveratrol on skin wound healing, scarring, and aging. Int Wound J, 19(1), 9-28. https://doi.org/10.1111/iwj.13601
• Hosokawa, R., Nonaka, K., Morifuji, M., Shum, L., & Ohishi, M. (2003). TGF-beta 3 decreases type I collagen and scarring after labioplasty. J Dent Res, 82(7), 558-564. https://doi.org/10.1177/154405910308200714
• Huang, C., Dong, L., Zhao, B., Lu, Y., Huang, S., Yuan, Z., Luo, G., Xu, Y., & Qian, W. (2022). Anti-inflammatory hydrogel dressings and skin wound healing. Clin Transl Med, 12(11), e1094. https://doi.org/10.1002/ctm2.1094
• Huang, X., Sun, J., Chen, G., Niu, C., Wang, Y., Zhao, C., Sun, J., Huang, H., Huang, S., Liang, Y., Shen, Y., Cong, W., Jin, L., & Zhu, Z. (2019). Resveratrol Promotes Diabetic Wound Healing via SIRT1-FOXO1-c-Myc Signaling Pathway-Mediated Angiogenesis. Front Pharmacol, 10, 421. https://doi.org/10.3389/fphar.2019.00421
• Jagiello, K., Uchanska, O., Matyja, K., Jackowski, M., Wiatrak, B., Kubasiewicz-Ross, P., & Karuga-Kuzniewska, E. (2023). Supporting the Wound Healing Process-Curcumin, Resveratrol and Baicalin in In Vitro Wound Healing Studies. Pharmaceuticals (Basel), 16(1). https://doi.org/10.3390/ph16010082
• Kant, V., Gopal, A., Kumar, D., Pathak, N. N., Ram, M., Jangir, B. L., Tandan, S. K., & Kumar, D. (2015). Curcumin-induced angiogenesis hastens wound healing in diabetic rats. J Surg Res, 193(2), 978-988. https://doi.org/10.1016/j.jss.2014.10.019
• Kasuya, A., & Tokura, Y. (2014). Attempts to accelerate wound healing. J Dermatol Sci, 76(3), 169-172. https://doi.org/10.1016/j.jdermsci.2014.11.001
• Moore, R. A., Straube, S., Wiffen, P. J., Derry, S., & McQuay, H. J. (2009). Pregabalin for acute and chronic pain in adults. Cochrane Database Syst Rev(3), CD007076. https://doi.org/10.1002/14651858.CD007076.pub2
• Nogueira, B. C. F., Campos, A. K., Alves, R. S., Sarandy, M. M., Novaes, R. M. D., Esposito, D., & Goncalves, R. V. (2020). What Is the Impact of Depletion of Immunoregulatory Genes on Wound Healing? A Systematic Review of Preclinical Evidence. Oxidative Medicine and Cellular Longevity, 2020. https://doi.org/Artn 8862953 10.1155/2020/8862953
• Ozdemir, K. G., Yilmaz, H., & Yilmaz, S. (2009). In vitro evaluation of cytotoxicity of soft lining materials on L929 cells by MTT assay. J Biomed Mater Res B Appl Biomater, 90(1), 82-86. https://doi.org/10.1002/jbm.b.31256
• Plikus, M. V., Guerrero-Juarez, C. F., Ito, M., Li, Y. R., Dedhia, P. H., Zheng, Y., Shao, M., Gay, D. L., Ramos, R., Hsi, T. C., Oh, J. W., Wang, X., Ramirez, A., Konopelski, S. E., Elzein, A., Wang, A., Supapannachart, R. J., Lee, H. L., Lim, C. H., . . . Cotsarelis, G. (2017). Regeneration of fat cells from myofibroblasts during wound healing. Science, 355(6326), 748-752. https://doi.org/10.1126/science.aai8792
• Ronnov-Jessen, L., & Petersen, O. W. (1993). Induction of alpha-smooth muscle actin by transforming growth factor-beta 1 in quiescent human breast gland fibroblasts. Implications for myofibroblast generation in breast neoplasia. Lab Invest, 68(6), 696-707. https://www.ncbi.nlm.nih.gov/pubmed/8515656
• Salat, K., Gdula-Argasinska, J., Malikowska, N., Podkowa, A., Lipkowska, A., & Librowski, T. (2016). Effect of pregabalin on contextual memory deficits and inflammatory state-related protein expression in streptozotocin-induced diabetic mice. Naunyn Schmiedebergs Arch Pharmacol, 389(6), 613-623. https://doi.org/10.1007/s00210-016-1230-x
• Seo, G. Y., Lim, Y., Koh, D., Huh, J. S., Hyun, C., Kim, Y. M., & Cho, M. (2017). TMF and glycitin act synergistically on keratinocytes and fibroblasts to promote wound healing and anti-scarring activity. Exp Mol Med, 49(3), e302. https://doi.org/10.1038/emm.2016.167 Sinha, M., Gautam, L., Shukla, P. K., Kaur, P., Sharma, S., & Singh, T. P. (2013). Current perspectives in NSAID-induced gastropathy. Mediators Inflamm, 2013, 258209. https://doi.org/10.1155/2013/258209
• Sirin, D. Y., & Karaarslan, N. (2018). Evaluation of the effects of pregabalin on chondrocyte proliferation and CHAD, HIF-1alpha, and COL2A1 gene expression. Arch Med Sci, 14(6), 1340-1347. https://doi.org/10.5114/aoms.2018.73134
• Su, W. H., Cheng, M. H., Lee, W. L., Tsou, T. S., Chang, W. H., Chen, C. S., & Wang, P. H. (2010). Nonsteroidal anti-inflammatory drugs for wounds: pain relief or excessive scar formation? Mediators Inflamm, 2010, 413238. https://doi.org/10.1155/2010/413238
• Takeo, M., Lee, W., & Ito, M. (2015). Wound Healing and Skin Regeneration. Cold Spring Harbor Perspectives in Medicine, 5(1). https://doi.org/ARTN a023267 10.1101/cshperspect.a023267
• Teplicki, E., Ma, Q., Castillo, D. E., Zarei, M., Hustad, A. P., Chen, J., & Li, J. (2018). The Effects of Aloe vera on Wound Healing in Cell Proliferation, Migration, and Viability. Wounds, 30(9), 263-268. https://www.ncbi.nlm.nih.gov/pubmed/30256753
• Tsai, H. W., Wang, P. H., & Tsui, K. H. (2018). Mesenchymal stem cell in wound healing and regeneration. J Chin Med Assoc, 81(3), 223-224. https://doi.org/10.1016/j.jcma.2017.06.011
• Wagener, N., Di Fazio, P., Boker, K. O., & Matziolis, G. (2022). Osteogenic Effect of Pregabalin in Human Primary Mesenchymal Stem Cells, Osteoblasts, and Osteosarcoma Cells. Life-Basel, 12(4). https://doi.org/ARTN 496 10.3390/life12040496
• Zhang, K., Lu, J., Mori, T., Smith-Powell, L., Synold, T. W., Chen, S., & Wen, W. (2011). Baicalin increases VEGF expression and angiogenesis by activating the ERRalpha/PGC-1alpha pathway. Cardiovasc Res, 89(2), 426-435. https://doi.org/10.1093/cvr/cvq296
• Zhang, X., Kang, X., Jin, L., Bai, J., Liu, W., & Wang, Z. (2018). Stimulation of wound healing using bioinspired hydrogels with basic fibroblast growth factor (bFGF). Int J Nanomedicine, 13, 3897-3906. https://doi.org/10.2147/IJN.S168998